Novel compounds

Abstract

The present invention relates to novel coagulation factor FVIIa polypeptides.

1 . A Factor VII (“FVII”) variant that is (a) free of GLA residues and (b) comprises an amino acid sequence that differs from human Factor VII (SEQ ID NO:1) by one or more substitutions of amino acid residues relative to SEQ ID NO:1 in one or more positions selected from K157, V158, E296, M298, L305, M306, D309, S314, D334, S336, K337, and F374 of SEQ ID NO:1.  
     
     
         2 . The Factor VII variant according to  claim 1 , wherein K157 is substituted.  
     
     
         3 . The Factor VII variant according to  claim 1 , wherein V158 is substituted.  
     
     
         4 . The Factor VII variant according to  claim 1 , wherein E296 is substituted.  
     
     
         5 . The Factor VII variant according to  claim 1 , wherein M298 is substituted.  
     
     
         6 . The Factor VII variant according to  claim 1 , wherein L305 is substituted.  
     
     
         7 . The Factor VII variant according to  claim 1 , wherein M306 is substituted.  
     
     
         8 . The Factor VII variant according to  claim 1 , wherein D309 is substituted.  
     
     
         9 . The Factor VII variant according to  claim 1 , wherein S314 is substituted.  
     
     
         10 . The Factor VII variant according to  claim 1  wherein D334 is substituted.  
     
     
         11 . The Factor VII variant according to  claim 1 , wherein S336 is substituted.  
     
     
         12 . The Factor VII variant according to  claim 1 , wherein K337 is substituted.  
     
     
         13 . The Factor VII variant according to  claim 1  wherein F374 is substituted.  
     
     
         14 . The Factor VII variant according to  claim 1 , wherein at least one additional amino acid residue in the protease domain is substituted relative to human Factor VII.  
     
     
         15 . The Factor VII variant according to  claim 14 , wherein at least one amino acid corresponding to an amino acid at a position selected from 159-170 of SEQ ID NO:1 is substituted.  
     
     
         16 . The Factor VII variant according to  claim 14 , wherein at least one amino acid corresponding to an amino acid at a position selected from 290-304 of SEQ ID NO:1 has been replaced with any other amino acid.  
     
     
         17 . The Factor VII variant according to  claim 16 , wherein R304 is replaced by an amino acid selected from the group consisting of Tyr, Phe, Leu, and Met.  
     
     
         18 . The Factor VII variant according to  claim 14 , wherein at least one amino acid corresponding to an amino acid at a position selected from 307-312 of SEQ ID NO:1 is substituted.  
     
     
         19 . The Factor VII variant according to  claim 14 , wherein at least one amino acid corresponding to an amino acid at a position selected from 330-339 of SEQ ID NO:1 is substituted.  
     
     
         20 . The Factor VII variant according to  claim 14 , wherein A274 is substituted.  
     
     
         21 . The Factor VII variant according to  claim 20 , wherein A274 has been replaced by an amino acid selected from the group consisting of Met, Leu, Lys, and Arg.  
     
     
         22 . The Factor VII variant according to  claim 2 , wherein K1157 is replaced by an amino acid selected from the group consisting of Gly, Val, Ser, Thr, Asn, Gin, Asp, and Glu.  
     
     
         23 . The Factor VII variant according to  claim 3 , wherein V1158 is replaced by an amino acid selected from the group consisting of Ser, Thr, Asn, Gln, Asp, and Glu.  
     
     
         24 . The Factor VII variant according to  claim 4 , wherein E296 is replaced by an amino acid selected from the group consisting of Arg, Lys, Ile, Leu and Val.  
     
     
         25 . The Factor VII variant according to  claim 5 , wherein M298 is replaced by an amino acid selected from the group consisting of Lys, Arg, Gln, and Asn.  
     
     
         26 . The Factor VII variant according to  claim 6 , wherein L305 is replaced by an amino acid selected from the group consisting of Val, Tyr and Ile.  
     
     
         27 . The Factor VII variant according to  claim 7 , wherein M306 is replaced by an amino acid selected from the group consisting of Asp, and Asn.  
     
     
         28 . The Factor VII variant according to  claim 8 , wherein D309 is replaced by an amino acid selected from the group consisting of Ser, and Thr.  
     
     
         29 . The Factor VII variant according to  claim 9 , wherein S314 is replaced by an amino acid selected from the group consisting of Gly, Lys, Gln and Glu.  
     
     
         30 . The Factor VII variant according to  claim 10 , wherein D334 is replaced by an amino acid selected from the group consisting of Gly, and Glu.  
     
     
         31 . The Factor VII variant according to  claim 11 , wherein S336 is replaced by an amino acid selected from the group consisting of Gly, and Glu.  
     
     
         32 . The Factor VII variant according to  claim 12 , wherein K337 is replaced by an amino acid selected from the group consisting of Ala, Gly, Val, Ser, Thr, Asn, Gln, Asp, and Glu.  
     
     
         33 . The Factor VII variant according to  claim 13 , wherein said F374 has been replaced by an amino acid selected from the group consisting of Pro, and Tyr.  
     
     
         34 . The Factor VII variant according to  claim 33 , wherein said F374 has been replaced by Tyr.  
     
     
         35 . The Factor VII variant according to  claim 1 , wherein the Factor VII variant lacks residues 1-38 of SEQ ID NO:1.  
     
     
         36 . The Factor VII variant according to  claim 35 , wherein the Factor VII variant lacks residues 1-44 of SEQ ID NO:1.  
     
     
         37 . The Factor VII variant according to  claim 36 , wherein the Factor VII variant lacks residues 1-152 of SEQ ID NO:1.  
     
     
         39 . The Factor VII variant according to  claim 1 , wherein the ratio between the proteolytic activity of the Factor VII variant and the proteolytic activity of human Factor VIIa is at least about 1.25.  
     
     
         40 . A polynucleotide comprising a sequence that encodes a Factor VII variant according to  claim 1 .  
     
     
         41 . A pharmaceutically acceptable composition comprising a FVII polypeptide that is essentially free of GLA residues and and, optionally, a pharmaceutically acceptable carrier.  
     
     
         42 . The composition of  claim 41 , wherien the FVII polypeptide is a FVII variant according to  claim 1 .  
     
     
         43 . The composition of  claim 41 , wherein the FVII polypeptide is a wild-type human FVII that is essentially free of GLA residues.  
     
     
         44 . A method for treating a bleeding disorder, treating a bleeding episode, or enhancing hemostasis in a subject comprising administering a therapeutically or prophylactically effective amount of a FVII polypeptide that is essentially free of GLA residues to a subject in need thereof.  
     
     
         45 . The method of  claim 44 , wherein the method comprises administering a therapeutically or prophylactically effective amount of a FVII variant according to  claim 1  to the subject.